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( Seung Hyeok Seok ),( Min Won Baek ),( Hui Young Lee ),( Dong Jae Kim ),( Yi Rang Na ),( Sung Hoon Park ),( Hyun Kyoung Lee ),( Doug Young Ryu ),( Jae Hak Park ) 한국동물실험대체법학회 2007 한국동물실험대체법학회 학술대회집 Vol.2007 No.1
We first established a transgenic zebrafish line carrying a plasmid (phAhRE-EGFP) for detecting developmental toxicities caused by aryl hydrocarbon receptor (AhR) binding toxicants. The plasmid contains the 5` human cytochrome P4501A1 (CYP1A1) upstream sequence, which AhR ligands target, and enhanced green fluorescent protein (EGFP) gene, an accurate and quick, inducible reporter. To evaluate this system, we treated embryos of wild-type and the transgenic fish with four well-known AhR binding toxicants: 3,3`,4,4`,5-pentachlorobiphenyl (PCB126), 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD), 3-methylcholanthrene (3-MC), and β-naphthoflavone (β-NF). These toxicants induced a dose-dependent increase in morphologic disruption, indicating toxicity at early life-stages. The transgenic zebrafish were sensitive enough to these toxicants to express the CYP1A1 regulated EGFP. Moreover, fluorescence-based quantification of the reporter gene was consistent with dose dependent toxicity of the four model toxicants. In conclusion, the stable transgenic line will allow for extremely rapid and reproducible toxicological profiling of early life-stage embryo development in both aquatic organisms and humans.
Seung-Hyeok Seok,Yi Rang Na,Daun Jung,Gyo Jeong Gu 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.10
Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had MHCIIlowF4/80high as well as CD11c+CD11bhighCD80-CD64+MerTK+ phenotypes. In contrast, GM-BMDCs had MHCIIhighF4/80low and CD11chighCD8α-CD11b+CD80+CD64-MerTKlow phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing TNF, IL-1, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.
The Acceptance or Rejection of Syngeneic Murine Tumor Transplantability in C57BL/6 Mice
Seung Hyeok Seok,Jong Hwan Park,Min Won Baek,Hui Young Lee,Dong Jae Kim,Jae Hak Park 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1
16.7% rejection of wild-type Lewis lung carcinoma (LLC) 1×10⁶ cells in syngeneicC57BL/6 mice by s.c, injection in the axillary region was observed, even 50.8% rejection with 1×10⁵ cells injection. To address this question, the well-characterized LLC model was chosen for this study. The growth of LLC in syngeneic C57BL/6 mice is accompanied by a weak and transient antitumor cytotoxic response that is later replaced by tumor-enhancing activity, which is correlated with the generation of suppressor cells in LLC-bearing mice. For the rejection of syngeneic tumor transplantation, ultimately "spreading" of the T cell immune response to novel antigenic epitopes needed. In summary, these in vivo transplant studies demonstrate that syngeneic tumor cell transplantation can potentiate the immune response.